Tailored Cyclodextrin Pore Blocker Protects Mammalian Cells from Clostridium difficile Binary Toxin CDT

Toxins

Ekaterina M. Nestorovich¹  Vladimir A. Karginov²  Carsten Schwan³  Klaus Aktories³  Holger Barth⁴  Maurice Roeder⁴ 

[1] Department of Biology, the Catholic University of America, Washington, DC 20064, USA;Innovative Biologics, Inc., 13455 Sunrise Valley Dr., Suite 200, Herndon, VA 20171, USA;Institute of Experimental and Clinical Pharmacology and Toxicology, University of Freiburg, 79104 Freiburg, Germany;Institute of Pharmacology and Toxicology, University of Ulm Medical Center,Albert-Einstein-Allee 11, 89081 Ulm, Germany;

关键词: cellular uptake;    Clostridium difficile CDT;    binary toxin;    membrane transport;    translocation pore;    pore blocker;    β-cyclodextrin;   

DOI  :  10.3390/toxins6072097

来源: DOAJ

【 摘 要 】

Some Clostridium difficile strains produce, in addition to toxins A and B, the binary toxin Clostridium difficile transferase (CDT), which ADP-ribosylates actin and may contribute to the hypervirulence of these strains. The separate binding and translocation component CDTb mediates transport of the enzyme component CDTa into mammalian target cells. CDTb binds to its receptor on the cell surface, CDTa assembles and CDTb/CDTa complexes are internalised. In acidic endosomes, CDTb mediates thedelivery of CDTa into the cytosol, most likely by forming a translocation pore in endosomal membranes. We demonstrate that a seven-fold symmetrical positively chargedβ-cyclodextrin derivative, per-6-S-(3-aminomethyl)benzylthio-β-cyclodextrin, which was developed earlier as a potent inhibitor of the translocation pores of related binary toxins of Bacillus anthracis, Clostridium botulinum and Clostridium perfringens, protects cells from intoxication with CDT. The pore blocker did not interfere with the CDTa-catalyzedADP-ribosylation of actin or toxin binding to Vero cells but inhibited the pH-dependent membrane translocation of CDTa into the cytosol. In conclusion, the cationicβ-cyclodextrin could serve as the lead compound in a development of novel pharmacological strategies against the CDT-producing strains of C. difficile.

【 授权许可】

Unknown